LOCUS       sequence-194378-        9600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            with mutated PAM2 motif in D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9600)
  AUTHORS   Chekulaeva M, Mathys H, Zipprich JT, Attig J, Colic M, Parker R, 
            Filipowicz W
  TITLE     miRNA repression involves GW182-mediated recruitment of CCR4-NOT 
            through conserved W-containing motifs.
  JOURNAL   Nat Struct Mol Biol. 2011 Oct 7;18(11):1218-26. doi: 
            10.1038/nsmb.2166.
  PUBMED    21984184
REFERENCE   2  (bases 1 to 9600)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nsmb"; 
            journalName: "Nat Struct Mol Biol"; date: "2011-10-7- 7"; volume: 
            "18"; issue: "11"; pages: "1218-26"
COMMENT     Sequence Label: pMCh-695
FEATURES             Location/Qualifiers
     source          1..9600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1746..1811
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1839..1865
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6065..6106
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6116..6133
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6156..6173)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6206..6340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6230..6249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6284..6303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6941..7529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7021..7040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7700..8491,8492..8560))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8323..8342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8561..8665)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8733..8751
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8815..9600,1..1725)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 tttcttggct tattcaaaaa gcaaagtaaa gccaaaaaag cgagatggca ataccaaatg
       61 cggcaaaacg gtagtggaag gaaaggggtg cggggcagcg gaaggaaggg tggggcgggg
      121 cgtggcgggg tctgtggctg ggcgcgacgt caccgacgtt ggagccactc ctttgaccat
      181 gtgtgcgtgt gtgtattatt cgtgtctcgc cactcgccgg ttgttttttt ctttttatgc
      241 tgcgctctct ctagcgccat ctcgcttacg catgctcaac gcaccgcatg ttgccgtttc
      301 cttttatgcg tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa
      361 aacgctaaaa taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg
      421 atagcaacaa aagcaacaac acaggtggct ttccctcttt cactttttgt ttgcaagccg
      481 cgtgcgagca agacggcacg accggcaaac gcaattacgc tgacaaagag cagacgaagt
      541 tttggcgaaa aacatcaagg cgcctgatac gaatgcattt gcaataacaa ttgcgatatt
      601 taatattgtt tatgaagctg tttgacttca aaacacacaa aaaaaaaaaa taaaacaaat
      661 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
      721 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
      781 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
      841 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
      901 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
      961 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1021 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1081 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1141 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     1201 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     1261 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     1321 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     1381 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     1441 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     1501 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     1561 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     1621 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     1681 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     1741 ccaacatgga cgcacaaaca cgacgacgtg agcgtcgcgc tgagaaacaa gctcaatgga
     1801 aagctgcaaa cggtggatct cctccaagct tgatggctta cccatacgat gttccagatt
     1861 acgctcctgc aggcatgcgt gaagcccttt tttcccaaga tggctggggc tgtcagcatg
     1921 ttaaccagga tactaattgg gaagttccca gttcgccaga accagccaat aaggatgcac
     1981 ccggtccacc aatgtggaag ccaagcatta acaatggtac tgatctttgg gagtccaatt
     2041 tgagaaacgg aggtcagccg gccgcacagc aagttccaaa gccgtcgtgg ggtcatacac
     2101 catcctctaa cttaggtgga acatggggtg aggacgacga tggcgccgat agtagtagtg
     2161 tgtggactgg aggagctgtt agcaacgcgg gatccggagc tgcagtggga gtaaaccaag
     2221 ccggagttaa tgtcggtcca ggcggtgttg tttcgtctgg cggacctcag tggggacaag
     2281 gtgtcgttgg cgtcggactt ggatcaactg gaggtaacgg gtcaagcaat ataactggat
     2341 cgtctggagt cgcaacaggt agtagcggaa actccagcaa cgctggtaac ggttggggag
     2401 accctcgtga aatacgccct ttgggagttg gtggctccat ggatattcga aatgttgaac
     2461 atcgcggcgg taacggttct ggagcaactt cgagcgatcc acgagacatt cgcatgatcg
     2521 atccgcgtga ccctattcga ggagatcccc gtggaatatc tggtcgtctt aatgggacct
     2581 ctgaaatgtg gggtcatcat ccacaaatgt cccataacca gttgcaaggt atcaacaaaa
     2641 tggttggtca aagtgtagca actgccagca ccagtgtcgg aacatctggc tcgggcatcg
     2701 gtcctggagg tcccggtcct agtacagtat caggcaatat cccaacacag tgggggcctg
     2761 ctcaaccggt aagcgttggt gtaagtggtc ccaaagacat gtcaaaacag ataagtggat
     2821 gggaggaacc atcaccaccg cctcagcgtc gcagtattcc taactacgat gatggtacat
     2881 cgttgtgggg tcagcaaact cgtgttcccg ctgcaagcgg tcactggaaa gacatgactg
     2941 attcgatagg tcgtagtagt catctcatgc gtggccaaag ccaaacggga ggtataggaa
     3001 tagccggcgt tggaaatagc aatgttccag tgggagccaa tccaagtaat cctataagca
     3061 gtgtagttgg acctcaagcc cggattccat ctgtgggcgg cgtacaacac aaaccagacg
     3121 gcggcgctat gtgggtgcat tccggcaatg taggtggcag aaataatgtt gctgctgtta
     3181 ctacttgggg agatgacact catagcgtta atgtcggcgc tcccagcagt ggcagtgtat
     3241 ccagcaacaa ttgggttgat gacaagtcca actcaacctt ggcacaaaac tcttggagcg
     3301 acccggcccc tgttggagtt agttggggca ataagcaaag caaaccgcca agcaatagtg
     3361 cttcatcagg ttggagcact gctgcgggcg tggtggatgg ggttgatcta ggatctgagt
     3421 ggaacacgca cggggggatt attggaaaat ctcagcaaca acaaaaacta gcgggactta
     3481 acgtgggaat ggtgaacgta attaacgcgg agatcattaa gcaaagcaag caatacagga
     3541 tccttgtcga gaacggcttt aaaaaggaag atgtagagcg ggcattagtg attgctaata
     3601 tgaacatcga agaggcagcc gatatgctcc gtgccaactc atccctatca atggatggtt
     3661 ggcgtcgaca tgatgagtcc cttggatctt atgccgacca caatagttca acaagcagcg
     3721 gtggatttgc tggtcgttac ccggtcaaca gtggacaacc ttcaatgtcc tttcctcata
     3781 ataaccttat gaataacatg ggaggtaccg ctgttactgg aggtaacaac aatacaaaca
     3841 tgacagcttt acaggtgcaa aagtatttaa atcaagggca acatggtgtc gctgttggac
     3901 cgcaagccgt tggtaattct tcagcagtat ctgtcggatt tggtcagaac acgtctaacg
     3961 cagcagtggc aggagcagcc tctgcaaata tagcagcaaa tacaaacaac caaccgtctg
     4021 gtcagcaaat tcgcatgcta ggccagcaaa ttcagttggc cattcatagt ggtttcatat
     4081 ctagtcagat attgactcaa ccgctaactc aaacaaccct taacctttta aaccaacttc
     4141 ttagcaatat taagcatctc caggctgcac agcaatccct tacccgcggg ggaaatgtca
     4201 atccaatggc agtgaatgtg gctatatcta aatacaagca gcaaatccag aatttacaga
     4261 accagataag tgcacaacag gctgtgtatg taaaacagca aaatatgcaa ccaacttcac
     4321 aacaacaaca gccccaacaa cagcaacttc cttctgttca tctaagtaac tcaggcaacg
     4381 actatttaag aggtcacgat gcaataaata atttgcaaag caacttttct gagctcaata
     4441 ttaataagcc aagtggatat caaggagcgt ccaatcaaca atcccgatta aatcagtgga
     4501 agcttccagt attagataag gagatcaact ctgacagtac ggaattttct cgtgccccag
     4561 gtgcaacgaa acaaaatttg acggccaaca caagcaacat aaactctttg ggtcttcaaa
     4621 acgatagtac atggtcaact ggacgcagta ttggtgacgg ttggcctgat ccctcatctg
     4681 ataacgagaa taaagactgg tctgttgctc agccaacttc agcagcaact gcttacactg
     4741 atctggtcca agcggcggag ccaggcaagc cagcggcggg ttcacagatc aaaagcatag
     4801 aagatgatcc cagcattgca ccaggaagcg ttgctagatc tccattgtct attaattcga
     4861 cgccaaaaga tgctgacata tttgccaata ccggtaaaaa ttcaccgact gatttaccgc
     4921 cactaagttt atcgtcgtct acatggagtt ttaatccaaa ccaaaattat ccgagtcaca
     4981 gttggtctga caatagtcaa caatgtaccg ccacttcgga gctttggaca agcccgctaa
     5041 ataaatcatc gtctcgaggt cccccgccag gattgactgc caattcaaat aaatctgcaa
     5101 atagtaatgc gtcaacgcca acaactatta ccggaggtgc gaatggatgg ttacagcctc
     5161 gaagtggcgg tgttcaaacc acaaacacta attggacagg tggtaacacc acttggggct
     5221 ccagttggtt gcttttgaaa aatctaacag cacagattga tggtcctact ttgcgtacac
     5281 tgtgtatgca gcatgggccc cttgtcagct ttcacccgta tttgaaccaa ggaattgcct
     5341 tatgtaaata tactactcgt gaggaggcga acaaggcgca aatggcgtta aacaactgtg
     5401 tcctcgccaa caccacaata tttgctgaat ctcccagcga gaacgaggtg caaagcatta
     5461 tgcagcactt accacaaact ccttcctcta caagctctag tggaactagt ggtggcaacg
     5521 tcggaggcgt cggcacttca gccaataatg caaacagtgg ttctgcagct tgtctgtccg
     5581 gaaacaatag cggcaacgga aacggcagcg cgagcggcgc cggcagcggc aacaatggca
     5641 acagtagctg caacaacagt gccgccgggg ggggcagcag cagcaacaac acgattacca
     5701 ctgtagcaaa ttcgaatctt gttggttcta gtggctctgt ctcaaattcc tctggcgtta
     5761 ctgctaactc tagtactgtt tctgtagtta gttgtacagc gagtgggaat tccataaatg
     5821 gggcaggtac tgcaaacagt tctggttcaa agagtagtgc aaacaattta gctagcggcc
     5881 agtctagcgc ttctaactta actaatagca ccaattcaac atggcgacaa actagccaaa
     5941 accaagctct tcaaagtcaa agcaggccat caggcagaga agctgacttt gattatatat
     6001 ctctcgttta ttccattgtt gatgattaag cggccgctcg agtctagagg gcccgcggtt
     6061 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     6121 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     6181 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     6241 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6301 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     6361 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     6421 caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc ttcaacccag
     6481 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     6541 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     6601 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     6661 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     6721 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     6781 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     6841 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc
     6901 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6961 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7021 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7081 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7141 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7201 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7261 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7321 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7381 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7441 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7501 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7981 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     8041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8281 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     8341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8581 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8641 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8701 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8761 ttcaagaatt aattctatat tctaaaaaca caaatgatac ttctaaaaaa aaatcatgaa
     8821 tggcatcaac tctgaatcaa atctttgcag atgcacctac ttctcatttc cactgtcaca
     8881 tcatttttcc agatctcgct gcctgttatg tggcccacaa accaagacac gttttatggc
     8941 cattaaagct ggctgatcgt cgccaaacac caaatacata atgaatatgt acacattcga
     9001 gaaagaagcg atcaaagaag cgtcttcggg cggagtagga gaatgcggag gagaaggaga
     9061 acgagctgat ctagtatctc tccacaatcc aatgccaact gaccaactgg ccatattcgg
     9121 agcaatttga agccaatttc catcgcctgg cgatcgctcc attcttggct atatgttttt
     9181 caccgttacc cggggccatt ttcaaagact cgtcggcaag ataagattgt gtcactcgct
     9241 gtctctcttc atttgtcgaa gaatgctgag gaatttcgcg atgacgtcgg cgagtatttt
     9301 gaagaatgag aataatttgt atttatacga aaatcagtta gtggaatttt ctacaaaaac
     9361 atgttatcta tagataattt tgttgcaaaa tatgttgact atgacaaaga ttgtatgtat
     9421 atacctttaa tgtattctca ttttcttatg tatttataat ggcaatgatg atactgatga
     9481 tattttaaga tgatgccaga ccaaaaggct tgaatttctg cgtcttttgc cgaacgcagt
     9541 gcatgtgcaa ttgttgtttt ttggaatatt caattttcgg actgtccgct ttgatttcag
//
